IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS) e-ISSN: 2278-3008, p-ISSN:2319-7676. Volume 10, Issue 4 Ver. II (Jul - Aug. 2015), PP 05-08 www.iosrjournals.org DOI: 10.9790/3008-10420508 www.iosrjournals.org 5 | Page Fabrication of Microfluidic Biosensor Minal B Engavale and Meghshree V Deshmukh Department of Microbiology, Ramnarian Ruia College, Matunga (E), Mumbai 400 092-India Abstract: Biosensors have been developed and used in a wide variety of ways like environmental monitoring of pollutants, detection of biological components in health care management etc. Fabrication of biosensors is not only a labour intensive process but also a very expensive one. The present study aims at developing a rapid, cost and labour effective, multi analytic microfluidic chip that can be used for the detection of markers responsible for any disease condition. One such marker uric acid, that is responsible for Gout a kidney disorder, was detected on the developed platform. The chip can also facilitate multi-analysis of a single sample on a miniaturized platform using flow through systems. The prepared device contains Sodium alginate beads with the entrapped Urease enzyme which detects the uric acid at Micron level. The entire device is established on small chip sized area (2 cm x 3cm) utilizing less volume of sample and reagents. The device is cost effective and rapid in its action. Key words: Microfluidics, PDMS, Biosensor, Urease. I. Introduction Biosensors have been developed and used in a wide variety of analytical settings including biomedical; health care; environmental monitoring; detection of biological, chemical, and toxic agents; physics; research and others. Several enzyme sensors have been developed using electrochemical, acoustic or optical systems. The importance of enzyme-based micro fluidic biosensors has increased considerably during the past decade due to high selectivity of the bio recognition element and the sensitivity of electrochemical signal transduction. Compared with conventional sensing techniques, distinctive advantages of using MFS for biomedicine include ultra-high sensitivity, higher throughput, in-situ monitoring and lower cost through cells, to small multi cellular organisms has explosively grown based on the advancement in micro fluidic system. Such technology also has the benefit of scaling the dimensions that enables a range of fundamental features to accompany system miniaturization such as reduced reagent consumption, high temporal resolution due to rapid mixing, high throughput, enhanced analytical performance, less waste, low unit cost, reduced energy consumption, and reduced dimensions when compared to macro scale techniques. There are various materials that can be used for fabrication one of the commonly used material is PDMS, PDMS device fabrication is one of the easiest methods for the rapid prototyping of micro fluidic devices. It is a flexible elastomeric polymer that is an excellent material for micro fluidic device fabrication (6, 5). Considering the above mentioned facts, it would be interesting and innovative to design new platforms. This research thus aims at addressing detection of markers responsible for any disease condition. The developed chip will help detect uric acid in the blood. This will be a basic platform which will involve immobilization of the enzymes in multi-channels and detection of the conversion products by chemical reagents. Microfluidics presents a very cheap, inexpensive and easiest way detecting the two on the same platform making it a multi enzyme micro fluidic based biosensor. Different technologies have been developed in recent years to produce multi analyte sensors (5). The development of large-scale biosensor arrays composed of highly miniaturized signal transducer elements, for example, enables the real-time parallel monitoring of species and is an important driving force in biosensor research. Said this, determination of analytes has been a great challenge. II. Materials And Methods 1. Extraction of Urease enzyme and its activity. Materials: 24 hours old grown culture of Urease producing organism ( Staphylococcus aureus isolated from urinal soil sample) in Urease broth, 50% Acetone. Method: For extracellular enzyme, 10 ml of the culture was taken into a sterile centrifuge tube and allowed to spin at 4 0 C at 5000 rpm for 20 minutes .The pellet was later discarded and the supernatant was taken in various ratios (1:2, 1:4, 1:5) with acetone. The mixture was stored in stationary conditions overnight at 4 o C.The pellet was separated out by centrifugation at 5000 rpm for 20 minutes at 4 o C the supernatant was discarded. To the pellet 10% of glycerol was added to solubilise the proteins and the solution was used for enzyme assay. (14)